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Total fibrin and fibrinogen degradation products in urine: a possible probe to detect illicit users of the physical-performance enhancer erythropoietin?

Erythropoietin (Epo) represents for some athletes the ultimate tool to gain an edge over their peer competitors. Underground information indicates that its usage is spreading at an epidemic pace since no analytical technique is yet available to detect its utilization. We hereby report observations obtained from analysis of urine specimens collected from top-level athletes after international-calibre competitions. Possible Epo misuse was evaluated by the measurement of urine total degradation products (TDPs), excretory fragments attributed by Sakakibara et al. to the fibrinolytic action of Epo. Markedly elevated urine TDP levels were measured in more than 13% of the 76 top-level athletes evaluated in this study. Analyses of urine specimens from a control hockey player group and from out-of-competition resting subjects indicate that the urine TDP content is not significantly influenced by exercise per se. Solid confirmation of TDP measurement as a sound probe to detect illicit Epo users should come from controlled studies with concomitant administration of Epo.

Adult↗

Estimation of dietary sodium intake in children.

To examine the utility of nocturnal urine collections in the estimation of dietary sodium intake in children, 29 families who agreed to lower their dietary sodium intake to less than 65 mEq/d were studied. A total of 58 adults and 95 children collected urine specimens weekly or biweekly before and after being instructed in a dietary sodium restriction regimen. Urine collections were fractionated into a diurnal and nocturnal portion. Sodium was measured directly, and estimated by measuring chloride excretion with a chloride titrator stick. Nocturnal chloride and 24-hour sodium excretion were highly correlated in adults (r = .75), and slightly less so (r = .56) in children. The slopes of the relationships were not different. Discriminant analysis showed that two or more chloride estimates permitted classification of both adults and children into the low sodium intake category with 100% accuracy, and into the normal sodium intake category with 75% accuracy. It was concluded that nocturnal chloride estimates are as useful in estimating compliance to a reduced sodium intake in children as in adults.

Adolescent↗

Hydrolysis of digoxin by acid.

Predictable hydrolysis of [3H]digoxin-12alpha occurred in vitro with incubation in HCl or gastric juice. Hydrolysis varied with pH, time, temperature and agitation. Digoxin, the bis- and mono-digitoxosides of digoxigenin and digoxigenin were separated by silica gel thin-layer chromatography using chloroform-ethyl acetate-glacial acetic acid (25:25:1 v/v) and were quantitated by liquid scintillation spectrometry. Hydrolysis with incubation at 37 degrees and pH 3 for 90 min was minimal, but increased with increasing acidity until greater than 70% was hydrolysed at pH 1-2 after 30 min and greater than 96% after 90 min incubation. At pH 0-9, 87% was hydrolysed after 30 min. In vitro hydrolysis in gastric fluid was slightly less than in HCl at the same pH. A volunteer was given 150 muCi[3H]digoxin-12alpha by nasogastric tube during a pentagastrin infusion when gastric pH was 0-94. He remained on his left side and samples were aspirated at intervals and immediately neutralized. Ethanol-chloroform 50-50 (v/v) extracts of the gastric fluid aspirated after 90 min and of all the urine specimens collected for 5 days were applied to a DEAE Sephadex LH-20 column. The radioactivity appeared in a single peak as digoxigenin in the 90 min gastric aspirate and in all urine specimens. Extensive intragastric hydrolysis of digoxin may occur under conditions of maximum acid output.

Adult↗

[The significance of determination of urinary type IV collagen concentrations from a random urine collection in patients with non-insulin dependent diabetes mellitus].

Recently, we have developed a highly sensitive assay system for quantitative measurement of urinary type IV collagen. To obtain a basic evaluation of the assay system we used urine specimens, collected randomly and studied the relation to diabetic nephropathy in patients with non-insulin dependent diabetes mellitus (NIDDM). It was shown that this assay system could measure the samples without concentrating the urine. Urinary sediments which were reported to interfere with the assay could be removed by adding Tris-buffer. In the clinical studies, urinary type IV collagen concentrations were significantly higher in patients with NIDDM regardless of the presence or the absence of albumin. Cases with abnormal values of urinary type IV collagen in the normal albuminuria group were 41.6%, while those with abnormal values of urinary transferrin excretion were 31.0%. Furthermore, among the normal albuminuria group, glycosylated hemoglobin (HbA1c) levels were higher in the groups of abnormal values of urinary type IV collagen than those within the reference values. These results obtained herein suggest that urinary type IV collagen could be a useful marker for the early stage of diabetic nephropathy.

Adult↗

Evaluation of suprapubic bladder aspiration for detection of group B streptococcal antigen by latex agglutination in neonatal urine.

Our objective was to determine whether urine collection by suprapubic bladder aspiration (SBA) improves the specificity of the group B streptococcal (GBS) latex agglutination (LA) test by avoiding contamination of urine with GBS from perineal and rectal colonization that can result in a positive LA test in an uninfected infant when the urine is collected by bag. Part 1 consists of a retrospective review of the medical records of 113 infants who had urine collected by SBA for GBS LA testing as part of evaluation for possible sepsis. The sensitivity and specificity of the urine LA test was assessed by comparing it with blood culture results. In part 2, a prospective analysis was performed of 19 newborns who had rectal and vaginal/penile cultures as well as urine by SBA and bag for GBS cultures and LA. Results of LA testing on urine collected by both of these methods were compared with results of urine, perineal, and rectal cultures. In the retrospective review of GBS LA testing performed on 113 consecutive urine specimens collected by SBA from neonates being evaluated for suspected sepsis, the sensitivity and specificity were 67% and 89%, respectively, when compared with blood culture results. Twelve infants who had a positive LA test result but a sterile blood culture (BC-,LA+) were compared with 95 infants with both blood cultures and urine LA tests negative for GBS (BC-, LA-). BC-, LA+ infants were more likely than those with BC-, LA- to have an immature to total neutrophil (I/T) ratio > or = 0.16 at 12 and 24 hours (p = 0.04 and 0.02, respectively). In the prospective study, we found that a positive GBS LA test can be due to perineal contamination and possibly to gastrointestinal absorption of GBS antigen. No false positive LA test results occurred on urine obtained by SBA; however, use of this method failed to detect the one infant with GBS bacteremia. Because of suboptimal sensitivity and specificity, use of the GBS LA test on urine obtained either by SBA or bag cannot be recommended for diagnosis of early onset GBS disease.

Antigens, Bacterial↗

An audit of the effectiveness of amphetamine isomer ratios as a marker of Dexedrine compliance in amphetamine abusers.

The effectiveness of using amphetamine isomer ratios to determine individual compliance with a Dexedrine (dexamphetamine, Celltech, Slough, UK) treatment regimen for the maintenance and detoxification from amphetamine abuse has been audited. The calculation of l/d-amphetamine isomer ratios in urine has been shown previously to provide an accurate method for determining the compliance of subjects prescribed dexamphetamine. This audit was performed on the results obtained subsequent to the analysis of 369 urine specimens collected from 87 subjects (26 female, 61 male), all prescribed dexamphetamine collected from January 1999 to December 2002. The analysis was performed using a chiral derivatizing agent in conjunction with a gas chromatograph fitted with a mass spectrometer. It was found that only 245 specimens (66%) were deemed to be from compliant subjects. Of the remainder, 60 (16%) were collected when subjects were apparently supplementing their dexamphetamine prescription with illicit amphetamine, and 66 (18%) were collected when subjects were apparently using primarily illicit amphetamine. In summary, a sensitive and specific technique has been developed and applied to the analysis of urine specimens obtained from those prescribed dexamphetamine to facilitate the monitoring of their compliance. It is hoped that this technique may continue to be used to audit further the treatment in these individuals in the future to optimize their management.

Adult↗

Monitoring STI prevalence using telephone surveys and mailed urine specimens: a pilot test.

OBJECTIVES: This pilot test assessed the feasibility of a cost effective population based approach to STI monitoring using automated telephone interviews, urine specimen collection kits sent out and returned by US Postal Service mail, and monetary incentives to motivate participation. METHODS: 100 residents of Baltimore, MD, USA, completed an automated telephone survey and agreed to mail in a urine specimen to be tested for chlamydia and gonorrhoea. Participants were paid dollar 10 for completing the survey and dollar 40 for mailing the specimen. RESULTS: 86% of survey participants mailed in a urine specimen for testing. CONCLUSIONS: Automated telephone surveys linked with testing of mailed-in urine specimens may be a feasible lower cost (relative to household surveys) method of estimating infection prevalences in a population.

Adolescent↗

Urine detection of survivin is a sensitive marker for the noninvasive diagnosis of bladder cancer.

PURPOSE: In a preliminary study urine detection of survivin, an integrator of cell death and mitosis, accurately detected bladder cancer. The objectives of this study were to confirm these findings in a large cohort of subjects undergoing cystoscopy, to assess the diagnostic performance of urine survivin and to test whether evaluation of urine survivin adds independent value to urine NMP22 (Matritech, Cambridge, Massachusetts) and cytology for the detection of bladder cancer. MATERIALS AND METHODS: Urine survivin was measured using a Bio-Dot microfiltration detection system (Bio-Rad, Hercules, California) in voided urine specimens collected before cystoscopy in 117 cases and 92 controls. Bladder washout samples for cytology were collected in 174 subjects. Urine levels of NMP22 were measured using a commercially available enzyme-linked immunosorbent assay. RESULTS: Higher levels of urine survivin were associated with an increased risk of bladder cancer (p <0.001) and tumors of higher grade (p = 0.037), but not with invasive stage, after adjustment for the effects of urine cytology, NMP22 and age. The sensitivity, specificity, and positive and negative predictive values of survivin for the diagnosis of bladder cancer (64%, 93%, 92% and 67%, respectively), are superior to those of NMP22 and cytology. Survivin had the highest specificity and positive predictive value for the detection of bladder cancer across each tumor stage and grade. CONCLUSIONS: Urine survivin was a strong, independent predictor of the presence of bladder cancer and higher tumor grade. Urine detection of survivin is an accurate diagnostic test for bladder cancer that retains its efficiency regardless of cancer stage and grade.

Adult↗

Urinary antimony in infancy.

OBJECTIVE: To determine whether antimony may be detected in the urine during infancy and early childhood and its association with passive exposure to tobacco smoke, as assessed by urinary cotinine. DESIGN: Analysis of spare aliquots of urine collected from infants participating in studies of respiratory function and passive smoking. Urinary antimony was assayed using inductively coupled plasma mass spectroscopy in 201 urine specimens collected at different ages throughout the first two years of life from 122 term and 26 preterm infants. Urinary cotinine was measured using gas liquid chromatography. MAIN OUTCOME MEASURE: Urinary antimony concentrations. RESULTS: Absolute antimony concentrations varied widely between infants, being below the laboratory detection limit of 0.02 microgram/l in 7% of samples, below 0.5 microgram/l in 90.5%, and above the reference value of 1 microgram/l reported for non-occupationally exposed UK populations in 4%. Creatinine standardised antimony values were unrelated to postnatal age or urinary cotinine concentrations and were highest in urine collected from preterm infants within 24 hours of birth (geometric mean (95% confidence interval): 2.3 ng/mg (1.5 to 3.4)). CONCLUSIONS: Although antimony is present at very low concentrations in urine during infancy and early childhood, the relevance to health is uncertain. The higher levels found in preterm infants may reflect prematurity or fetal assimilation of antimony. Tobacco is unlikely to be an important source of environmental exposure to antimony during infancy and early childhood.

Aging↗

Intrauterine diagnosis of cytomegalovirus and rubella infections by amniocentesis.

OBJECTIVE: To establish a correlation between the presence of cytomegalovirus (CMV) or rubella virus in amniotic fluid obtained through amniocentesis and fetal infection. DESIGN: Case series. SETTING: Five hospitals in the Montreal region. Virology testing was done at the Virology Research Centre, Institut Armand-Frappier, Laval, Que. PATIENTS: Thirteen pregnant women infected with CMV, 3 with rubella, their 15 babies and 2 fetuses. Twelve of the women with CMV infection were recruited from a prospective study of CMV infection in pregnancy. Infection in the other women was detected through routine laboratory diagnostic testing. INTERVENTION: Amniotic fluid samples were cultured for CMV and rubella virus. Congenital infection of the neonates was established through isolation of either virus from pharyngeal mucus and urine specimens collected during the first 3 days of life or from fetal tissue if the pregnancy was terminated. MAIN RESULTS: CMV was cultured from the amniotic fluid of three of the CMV-infected women and from the pharyngeal mucus and urine specimens of their infants. Of the three women with rubella the amniotic fluid of one (who had a twin pregnancy) was positive for rubella virus. After the in-utero death of one fetus she underwent a therapeutic abortion of both. Examination of fetal tissue indicated that both fetuses had been infected with rubella virus. Each of the two other women with rubella gave birth to an uninfected, healthy infant. CONCLUSIONS: We found a strong correlation between the isolation of CMV or rubella virus from the amniotic fluid and the presence of congenital infection. This suggests that amniocentesis used to detect the presence of a virus is a useful method for the diagnosis of fetal infection.

Adult↗

Significance of tissue fragments in voided urine specimens.

OBJECTIVE: To determine the diagnostic significance of tissue fragments in voided urine specimens. STUDY DESIGN: We reviewed all the voided urine specimens collected and processed by the Millipore filter technique (Bedford, Massachusetts, U.S.A.) at our institution between January 1, 1997, and December 31, 1998, and the corresponding biopsies obtained within 120 days after urine examination. The type and number of tissue fragments were correlated with the histologic diagnosis and clinical features; the results were compared to those from a recently published study. RESULTS: Of the 2,553 voided urine specimens examined, 174 (7%) had corresponding biopsies. Cell groups (tissue fragments) consisting of either flat sheets or three-dimensional structures were significantly more common (57%) (chi 2 P < .005) in urine specimens with biopsies revealing urothelial malignancies than in negative biopsies (6%). Three-dimensional groups were also statistically more common in cases with invasive transitional cell carcinoma. Proper identification of tissue fragments was highly significant and correlated with urothelial neoplastic changes. CONCLUSION: The results of the present investigation using the Millipore filter technique for voided urine cytology processing, differed from those of a recently published study that employed cytocentrifugation. Tissue fragments, to be differentiated from groups, clumps or clusters that often result from centrifugation and other concentration artifacts, were strongly associated with urothelial neoplasia, uncommonly with nonneoplastic disease processes affecting the urinary tract but always with urothelial disease.

Adult↗

EMIT-d.a.u. monoclonal amphetamine/methamphetamine assay. I. Stereoselectivity and clinical evaluation.

The stereoselectivity, cross-reactivity and clinical performance of the EMIT-d.a.u. monoclonal amphetamine(A)/methamphetamine (MA) immunoassay (EM) were evaluated. The cut-off calibrator of the assay was 1000 ng/ml S(+)MA. Analysis of drug-added urines and 72 clinical specimens demonstrated a cut-off for S(+)-amphetamine of approximately 400 ng/ml. The stereoisomeric selectivity of the assay was determined in a concentration vs. response manner by adding pure S(+) or R(-)isomers of A and MA, to drug free urine. The EM assay demonstrated a high selectivity for S(+)-isomers with only one of 16 urine specimens collected following excessive use of nasal inhalers yielding a positive result. This specimen contained 6000 ng/ml R(-)MA. Five-hundred clinical urine specimens were simultaneously analyzed for A or MA by the EM and EMIT-d.a.u. polyclonal (EP) amphetamine assay with 131 positive results confirmed by GC/MS. In five specimens negative by EM while positive by EP, MA was present at concentrations below the 1000 ng/ml cut-off. Two ME false positive results were apparently caused by chlorpromazine (CPZ) metabolites. A study of other phenothiazines or their metabolites gave no false positive results. The possible cross reactivity of the EM assay was further studied for phenyl-isopropylamine analogs or drugs previously reported to react with the EP assay. The EM assay showed much less cross-reactivity than EP to all drugs tested.

Amphetamine↗

LCR testing for gonorrhoea and chlamydia in population surveys and other screenings of low prevalence populations: coping with decreased positive predictive value.

OBJECTIVE: Nucleic acid amplification tests have facilitated field based STD studies and increased screening activities. However, even with highly specific tests, the positive predictive value (PPV) of such tests may be lower than desirable in low prevalence populations. We estimated PPVs for a single LCR test in a population survey in which positive specimens were retested. METHODS: The Baltimore STD and Behavior Survey (BSBS) was a population based behavioural survey of adults which included collecting urine specimens to assess the prevalence of gonorrhoea and chlamydial infection. Gonorrhoea and chlamydial infection were diagnosed by ligase chain reaction (LCR). Nearly all positive results were retested by LCR. Because of cost considerations, negative results were not confirmed. Predicted curves for the PPV were calculated for a single testing assuming an LCR test sensitivity of 95%, and test specificities in the range 95.0%-99.9%, for disease prevalences between 1% and 10%. Positive specimens were retested to derive empirical estimates of the PPV of a positive result on a single LCR test. RESULTS: 579 participants age 18-35 provided urine specimens. 20 (3.5%) subjects initially tested positive for chlamydial infection, and 39 (6.7%) tested positive for gonococcal infection. If positive results on the repeat LCR are taken as confirmation of a "true" infection, the observed PPV for the first LCR testing was 89.5% for chlamydial infection and 83.3% for gonorrhoea. This is within the range of theoretical PPVs calculated from the assumed sensitivities and specificities of the LCR assays. CONCLUSIONS: Empirical performance of a single LCR testing approximated the theoretically predicted PPV in this field study. This result demonstrates the need to take account of the lower PPVs obtained when such tests are used in field studies or clinical screening of low prevalence populations. Repeat testing of specimens, preferably with a different assay (for example, polymerase chain reaction), and disclosure of the non-trivial potential for false positive test results would seem appropriate in all such studies.

Adolescent↗

The detection of myoglobin in urine and its application to the diagnosis of myocardial infarction.

An indirect haemaglutination method was developed for the immunochemical detection of myoglobin in human urine. Myoglobin was found in the urine of 84% of 44 patients with acute myocardial infarction. In contrast, it was detected in the urine of 2.7% of 147 control subjects and in none of a control group of 15 patients who had received intramuscular injections. In patients with myocardial infarction, myoglobin was detected in most but not all urine specimens collected between 10 and 50 hours after the onset of chest pain. The results suggest that the detection of myoglobin in urine may be a valuable addition to present tests for the diagnosis of acute myocardial infarction.

Acute Disease↗

Biological monitoring of workers after the application of insecticidal pyrethroids.

OBJECTIVES: Pyrethroids are applied as insecticides throughout the world. Human metabolism of pyrethroids results in urinary metabolites that are suitable for biological monitoring. The aim of the study was to evaluate individual exposure due to occupational application of pyrethroids as a precondition for the assessment of health risks. METHODS: Thirty-six workers who applied insecticides and other pesticides in Germany collected samples of their urine (24 h) after having used various pyrethroids (alpha-cypermethrin, cypermethrin, cyfluthrin, deltamethrin, tau-fluvalinate, permethrin, lambda-cyhalothrin) in agriculture, greenhouses or indoor pest control. Biological monitoring was carried out and metabolites were analysed in 61 urine samples by GC-MS: cis-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid ( cis-Cl(2)CA and trans-Cl(2)CA), cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid ( cis-Br(2)CA), 3-phenoxybenzoic acid (3-PBA) and 4-fluoro-3-phenoxybenzoic acid (FPBA). Forty-five urine specimens collected (24 h) from persons with no occupational exposure to pyrethroids served as controls. Concentrations were related to creatinine content and expressed as microgrammes per gramme creatinine. RESULTS: Control urine samples revealed a considerable background excretion of pyrethroid metabolites by the general population. The 95th percentile of the concentrations of Cl(2)CA and cis-Br(2)CA were 2.1 and 0.1 microg/g creatinine, respectively. FPBA was not detected in any control urine and was found in only one sample within the complete study. After occupational application of pyrethroids the highest concentrations of metabolites in urine samples were detected within the group of indoor pest-control operators. The maximum concentrations (median values) of Cl(2)CA, 3-PBA, and cis-Br(2)CA were 92.4 microg/g (1.8 microg/g), 57.5 microg/g (1.4 microg/g) and 1.1 microg/g (median below detection limit), respectively. Workers in greenhouses excreted metabolites with median concentrations as follows: 2.9 microg/g Cl(2)CA, 0.5 microg/g cis-Br(2)CA and 2.9 microg/g 3-PBA. Medians of the metabolite concentrations in specimens from agricultural workers were below the detection limit with regard to Cl(2)CA and cis-Br(2)CA, but the value was 0.6 microg/g for 3-PBA. Pest-control operators excreted significantly higher concentrations of Cl(2)CA and 3-PBA than workers in agriculture on a collective basis. Comparison of the excreted concentrations of metabolites with values of acceptable daily intake (ADI) of pyrethroids set by WHO revealed that the amount of pyrethroids that had been taken up during occupational application was not considerably higher than the ADI. CONCLUSIONS: As a consequence, we conclude that adverse health effects are not to be expected after workers' occupational exposure to pyrethroids in Germany, provided that the application is carried out properly. Good working practices need to be supported by adequate supervision with regard to occupational hygiene and medicine.

Agriculture↗

A research note: the outcome of GC/MS/MS confirmation of hair assays on 93 cannabinoid (+) cases.

This article reports the outcome of gas chromatography/tandem mass spectrometry confirmations for THC and carboxy-THC on 93 hair samples screened by RIA for cannabinoids. The samples were taken from probationers in Pinellas County, FL, who voluntarily provided the research staff with six hair and six urine specimens, collected at 1-month intervals. There were 40 samples that were RIA (+), urinalysis (-). Samples were selected which had cannabinoid (+) outcomes for hair, urine, or both. The THC and/or the carboxy-THC was (+) on confirmation. Of these 40 samples, 22 were (+) for both THC and carboxy-THC, 15 were (+) for THC but not carboxy-THC, and three were carboxy THC (+), but THC (-). Only one sample had a (+) RIA, but was (-) for both THC and carboxy-THC on confirmation. RIA detection of cannabinoids was confirmed in nearly all cases. Most cases that were RIA (-) but urine (+) were cannabinoid (+) when analyzed by GC/MS/MS.

Cannabinoids↗

Use of multiple nucleic acid amplification tests to define the infected-patient "gold standard" in clinical trials of new diagnostic tests for Chlamydia trachomatis infections.

Nucleic acid amplification tests (NAATs) can be used to define the infected-patient "gold standard" for the purpose of designing studies of the performance of Chlamydia trachomatis diagnostic tests. It is unclear how many test results run by different NAATs and what combinations of specimens comprise the best infected-patient gold standard. We approached this question with data from a large study of the performance of a new NAAT. Data were available from three endocervical swabs and a urine specimen collected from each of 1,412 women and tested by three different NAATs. Results from all three assays were used equally in a rotating fashion to define the infected-patient gold standard. Multiple different infected-patient gold standards for estimating swab and urine specimen sensitivity and specificity for one NAAT method were created by varying the number and combinations of swab and urine comparator results with two different NAATs, The effect of changing the infected-patient gold standard definition was determined by constructing receiver-operator-like curves with calculated sensitivities and specificities for each test. The one-positive-of-two-results or two-positive-of-two-results (same or two different assays) infected-patient gold standard definitions produced low sensitivity and low specificity estimates, respectively. If four comparator NAAT results were used, the any-three-positive-of-four-results definition or the at-least-one-specimen-positive-by-each-of-two-comparator-assays definition appeared to provide better combinations of sensitivity and specificity estimates. The any-two-positive-out-of-three-results definition resulted in estimates that were as good as produced with the former two definitions. This analytic approach provides a means of clearly visualizing the effects of changing NAAT-based infected-patient gold standards and should be helpful in designing future studies of new C. trachomatis diagnostic tests.

Cervix Uteri↗

Psilocin identified in a DUID investigation.

Psilocin was identified in a urine specimen collected during a routine driving under the influence of drugs investigation, the first for this laboratory. The subject did not exhibit any response to an automobile crash, indicating the he may have been unaware of the severity of the situation or his immediate surroundings. The urine specimen gave a positive result on a fluorescence polarization immunoassay amphetamine/methamphetamine assay, and psilocin was determined to have 1.3% cross-reactivity at 50 mg/L. Psilocin was confirmed by gas chromatography-mass spectrometry following an extraction for acidic, neutral, and basic drugs. Hydrolysis and derivatization techniques were not employed. The urine concentration of psilocin decreased rapidly, although the specimen was maintained at 4 degrees C.

Accidents, Traffic↗